http://experimentationlab.berkeley.edu/sites/default/files/AFMImages/butt_AFM.pdf

Force measurements with the atomic force microscope: Technique, interpretation and applications

Biological materials: Structure and mechanical properties

A method to determine the spring constant of a rectangular atomic force microscope cantilever is proposed that relies solely on the measurement of the resonant frequency and quality factor of the cantilever in fluid (typically air), and knowledge of its plan view dimensions. This method gives very good accuracy and improves upon the previous formulation by Sader et al. [Rev. Sci. Instrum. 66, 3789 (1995)] which, unlike the present method, requires knowledge of both the cantilever density and thickness.

/pdf/calibration-of-rectangular-atomic-force-microscope-290lry8thn.pdf

Calibration of rectangular atomic force microscope cantilevers

Dynamic atomic force microscopy methods

Change in cell stiffness is a new characteristic of cancer cells that affects the way they spread1,2. Despite several studies on architectural changes in cultured cell lines1,3, no ex vivo mechanical analyses of cancer cells obtained from patients have been reported. Using atomic force microscopy, we report the stiffness of live metastatic cancer cells taken from the body (pleural) fluids of patients with suspected lung, breast and pancreas cancer. Within the same sample, we find that the cell stiffness of metastatic cancer cells is more than 70% softer, with a standard deviation over five times narrower, than the benign cells that line the body cavity. Different cancer types were found to display a common stiffness. Our work shows that mechanical analysis can distinguish cancerous cells from normal ones even when they show similar shapes. These results show that nanomechanical analysis correlates well with immunohistochemical testing currently used for detecting cancer.

Nanomechanical analysis of cells from cancer patients

Tapping mode atomic force microscopy in liquids gives a substantial improvement in imaging quality and stability over standard contact mode. In tapping mode the probe‐sample separation is modulated as the probe scans over the sample. This modulation causes the probe to tap on the surface only at the extreme of each modulation cycle and therefore minimizes frictional forces that are present when the probe is constantly in contact with the surface. This imaging mode increases resolution and reduces sample damage on soft samples. For our initial experiments we used a tapping frequency of 17 kHz to image deoxyribonucleic acid plasmids on mica in water. When we imaged the same sample region with the same cantilever, the plasmids appeared 18 nm wide in contact mode and 5 nm in tapping mode.

Tapping mode atomic force microscopy in liquids

We have designed and tested a family of silicon nitride cantilevers ranging in length from 23 to 203 μm. For each, we measured the frequency spectrum of thermal motion in air and water. Spring constants derived from thermal motion data agreed fairly well with the added mass method; these and the resonant frequencies showed the expected increase with decreasing cantilever length. The effective cantilever density (calculated from the resonant frequencies) was 5.0 g/cm3, substantially affected by the mass of the reflective gold coating. In water, resonant frequencies were 2 to 5 times lower and damping was 9 to 24 times higher than in air. Thermal motion at the resonant frequency, a measure of noise in tapping mode atomic force microscopy, decreased about two orders of magnitude from the longest to the shortest cantilever. The advantages of the high resonant frequency and low noise of a short (30 μm) cantilever were demonstrated in tapping mode imaging of a protein sample in buffer. Low‐noise images were tak...

Short cantilevers for atomic force microscopy

We present experimental support for a model of abalone nacre growth that is based on mineral bridges between successive aragonite tablets rather than on heteroepitaxial nucleation. Interlamellar sheets of organic polymers delineate the aragonite tablets but allow the tablets to grow mineral bridges through pores in the sheets. Atomic force microscope images of interlamellar organic sheets from flat pearls made by Haliotis rufescens (red abalone; marine gastropod mollusk) reveal a fibrous core and holes of 5−50 nm in diameter. Scanning ion conductance microscopy shows that these holes are actually pores through the interlamellar sheets. With the help of statistical analysis we can associate the pore-to-pore spacings in the interlamellar sheets with the observed offsets of successive nacre tablets. These results, supplemented by AFM, SEM, and TEM images, support and extend the model of biofabrication of gastropod nacre which is based on mineral bridges between the aragonite tablets.

/pdf/does-abalone-nacre-form-by-heteroepitaxial-nucleation-or-by-329q6ri22t.pdf

Does Abalone Nacre Form by Heteroepitaxial Nucleation or by Growth through Mineral Bridges

Imaging adhesion forces and elasticity of lysozyme adsorbed on mica with the atomic force microscope

An atomic force microscope (AFM) design providing a focused spot of order 7 μm in diameter was used to analyze the motion of vibrating cantilevers in liquid. Picking an operating frequency for tapping mode AFM operation in liquid is complex because there is typically a large number of sharp peaks in the response spectrum of cantilever slope amplitude versus drive frequency. The response spectrum was found to be a product of the cantilever’s broad thermal noise spectrum and an underlying fluid drive spectrum containing the sharp peaks. The geometrical shape of transverse cantilever motion was qualitatively independent of the fluid drive spectrum and could be approximately reproduced by a simple theoretical model. The measurements performed give new insights into the behavior of cantilevers during tapping mode AFM operation in liquid.

http://www.bioforce.uni-tuebingen.de/pubs/Schaffer_1996_JAP.pdf

D. A. Walters

Papers

Tapping mode atomic force microscopy in liquids

Short cantilevers for atomic force microscopy

Does Abalone Nacre Form by Heteroepitaxial Nucleation or by Growth through Mineral Bridges

Imaging adhesion forces and elasticity of lysozyme adsorbed on mica with the atomic force microscope

Studies of vibrating atomic force microscope cantilevers in liquid